Recombinant Lactobacillus with decreased lipoteichoic acid to reduce inflammatory responses

Abstract

Methods and compositions for treating or preventing inflammatory disorders are provided. The compositions of the invention comprise a recombinant bacterium genetically modified to decrease the display of lipoteichoic acid on the cell surface. Methods of the invention comprise administering to a subject a recombinant bacterium modified to decrease the display of lipoteichoic acid on the cell surface. Administration of the recombinant bacterium promotes a desired therapeutic response. The recombinant bacterium may be administered in a single dose or series of doses. Methods of the invention find use in treating or preventing a variety of inflammatory disorders including, for example, treating or preventing inflammatory bowel disease, colitis, or Crohn's disease.

That which is claimed: 
     
       1. A recombinant or isolated bacterium having been genetically modified to decrease the display of lipoteichoic acid (LTA) on the surface of said bacterium, wherein the genetic modification comprises deletion or substitution of at least one base pair such that the expression of at least one polynucleotide encoding a phosphoglycerol transferase is decreased,
 wherein said polynucleotide encoding a phosphoglycerol transferase comprises a nucleotide sequence having at least 85% sequence identity to the nucleic acid sequence as set forth in SEQ ID NO:1; and 
 wherein decreasing the expression of the at least one polynucleotide encoding a phosphoglycerol transferase decreases the display of LTA on the surface of the bacterium. 
 
     
     
       2. The recombinant or isolated bacterium of  claim 1 , wherein said polynucleotide encoding a phosphoglycerol transferase comprises a nucleotide sequence having at least 90% sequence identity to the nucleic acid sequence as set forth in SEQ ID NO:1. 
     
     
       3. The recombinant or isolated bacterium of  claim 1 , wherein said recombinant or isolated bacterium is a probiotic bacterium. 
     
     
       4. The recombinant or isolated bacterium of  claim 3 , wherein said probiotic bacterium is a lactic acid bacterium. 
     
     
       5. The recombinant or isolated bacterium of  claim 4 , wherein said lactic acid bacterium is a  Lactobacillus.    
     
     
       6. The recombinant bacterium of  claim 5 , wherein said  Lactobacillus  is  Lactobacillus acidophilus.    
     
     
       7. The recombinant or isolated bacterium of  claim 6 , wherein said genetic modification was performed on  Lactobacillus acidophilus  NCFM. 
     
     
       8. The recombinant or isolated bacterium of  claim 6 , wherein said  Lactobacillus acidophilus  is  Lactobacillus acidophilus  NCK2025, deposited under ATCC accession number PTA-11587. 
     
     
       9. A method of making a recombinant or isolated bacterium, said method comprising genetically modifying a bacterium to decrease the display of lipoteichoic acid (LTA) on the surface of said bacterium,
 wherein the genetic modification comprises deletion or substitution of at least one base pair such that the expression of at least one polynucleotide encoding a phosphoglycerol transferase is decreased, 
 wherein said polynucleotide encoding a phosphoglycerol transferase comprises a nucleotide sequence having at least 85% sequence identity to the nucleic acid sequence as set forth in SEQ ID NO:1; and 
 wherein decreasing the expression of the at least one polynucleotide encoding a phosphoglycerol transferase decreases the display of LTA on the surface of the bacterium. 
 
     
     
       10. The method of  claim 9 , wherein said recombinant or isolated bacterium has been genetically modified to decrease the expression of a polynucleotide comprising a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence as set forth in SEQ ID NO:1. 
     
     
       11. The method of  claim 9 , wherein said recombinant bacterium is a probiotic bacterium. 
     
     
       12. The method of  claim 11 , wherein said probiotic bacterium is a lactic acid bacterium. 
     
     
       13. A pharmaceutical composition comprising the recombinant or isolated bacterium according to  claim 1 . 
     
     
       14. The recombinant or isolated bacterium of  claim 1 , wherein said polynucleotide encoding a phosphoglycerol transferase comprises the nucleotide sequence as set forth in SEQ ID NO: 1. 
     
     
       15. The recombinant or isolated bacterium of  claim 1 , wherein said deletion or substitution comprises a deletion or substitution of at least one base pair of said polynucleotide encoding a phosphoglycerol transferase. 
     
     
       16. The method of  claim 9 , wherein said polynucleotide encoding a phosphoglycerol transferase comprises the nucleotide sequence as set forth in SEQ ID NO: 1. 
     
     
       17. The method of  claim 9 , wherein said deletion or substitution comprises a deletion or substitution of at least one base pair of said polynucleotide encoding a phosphoglycerol transferase.